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-基本情報
登録情報 | データベース: SASBDB / ID: SASDFM5 |
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試料 | Mutant 2-amino-3-carboxymuconate 6-semialdehyde decarboxylase, H110A tetramer, at pH 8.5
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機能・相同性 | 機能・相同性情報 secondary metabolic process / : / : / carboxy-lyase activity / hydrolase activity / metal ion binding / cytosol 類似検索 - 分子機能 |
生物種 | Pseudomonas fluorescens (蛍光菌) |
引用 | ジャーナル: J Biol Chem / 年: 2019 タイトル: Quaternary structure of α-amino-β-carboxymuconate-ϵ-semialdehyde decarboxylase (ACMSD) controls its activity. 著者: Yu Yang / Ian Davis / Tsutomu Matsui / Ivan Rubalcava / Aimin Liu / 要旨: α-Amino-β-carboxymuconate-ϵ-semialdehyde decarboxylase (ACMSD) plays an important role in l-tryptophan degradation via the kynurenine pathway. ACMSD forms a homodimer and is functionally inactive ...α-Amino-β-carboxymuconate-ϵ-semialdehyde decarboxylase (ACMSD) plays an important role in l-tryptophan degradation via the kynurenine pathway. ACMSD forms a homodimer and is functionally inactive as a monomer because its catalytic assembly requires an arginine residue from a neighboring subunit. However, how the oligomeric state and self-association of ACMSD are controlled in solution remains unexplored. Here, we demonstrate that ACMSD from can self-assemble into homodimer, tetramer, and higher-order structures. Using size-exclusion chromatography coupled with small-angle X-ray scattering (SEC-SAXS) analysis, we investigated the ACMSD tetramer structure, and fitting the SAXS data with X-ray crystal structures of the monomeric component, we could generate a pseudo-atomic structure of the tetramer. This analysis revealed a tetramer model of ACMSD as a head-on dimer of dimers. We observed that the tetramer is catalytically more active than the dimer and is in equilibrium with the monomer and dimer. Substituting a critical residue of the dimer-dimer interface, His-110, altered the tetramer dissociation profile by increasing the higher-order oligomer portion in solution without changing the X-ray crystal structure. ACMSD self-association was affected by pH, ionic strength, and other electrostatic interactions. Alignment of ACMSD sequences revealed that His-110 is highly conserved in a few bacteria that utilize nitrobenzoic acid as a sole source of carbon and energy, suggesting a dedicated functional role of ACMSD's self-assembly into the tetrameric and higher-order structures. These results indicate that the dynamic oligomerization status potentially regulates ACMSD activity and that SEC-SAXS coupled with X-ray crystallography is a powerful tool for studying protein self-association. |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-モデル
モデル #2961 | タイプ: mix / ダミー原子の半径: 1.90 A / カイ2乗値: 0.250 / P-value: 0.063313 Omokage検索でこの集合体の類似形状データを探す (詳細) |
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-試料
試料 | 名称: Mutant 2-amino-3-carboxymuconate 6-semialdehyde decarboxylase, H110A tetramer, at pH 8.5 |
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バッファ | 名称: 50 mM Tris, 5 mM DTT / pH: 8.5 |
要素 #1627 | 名称: ACMSD / タイプ: protein 記述: 2-amino-3-carboxymuconate 6-semialdehyde decarboxylase 分子量: 39.661 / 分子数: 4 / 由来: Pseudomonas fluorescens / 参照: UniProt: Q83V25 配列: MGHHHHHHHH HHSSGHIEGR HMKKPRIDMH SHFFPRISEQ EAAKFDANHA PWLQVSAKGD TGSIMMGKNN FRPVYQALWD PAFRIEEMDA QGVDVQVTCA TPVMFGYTWE ANKAAQWAER MNDFALEFAA HNPQRIKVLA QVPLQDLDLA CKEASRAVAA GHLGIQIGNH ...配列: MGHHHHHHHH HHSSGHIEGR HMKKPRIDMH SHFFPRISEQ EAAKFDANHA PWLQVSAKGD TGSIMMGKNN FRPVYQALWD PAFRIEEMDA QGVDVQVTCA TPVMFGYTWE ANKAAQWAER MNDFALEFAA HNPQRIKVLA QVPLQDLDLA CKEASRAVAA GHLGIQIGNH LGDKDLDDAT LEAFLTHCAN EDIPILVHPW DMMGGQRMKK WMLPWLVAMP AETQLAILSL ILSGAFERIP KSLKICFGHG GGSFAFLLGR VDNAWRHRDI VREDCPRPPS EYVDRFFVDS AVFNPGALEL LVSVMGEDRV MLGSDYPFPL GEQKIGGLVL SSNLGESAKD KIISGNASKF FNINV |
-実験情報
ビーム | 設備名称: Stanford Synchrotron Radiation Lightsource (SSRL) BL4-2 地域: Menlo Park, CA / 国: USA / 線源: X-ray synchrotron / 波長: 0.1228 Å / スペクトロメータ・検出器間距離: 2.5 mm | |||||||||||||||||||||||||||||||||
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検出器 | 名称: Pilatus3 X 1M / Pixsize x: 0.172 mm | |||||||||||||||||||||||||||||||||
スキャン | タイトル: Mutant 2-amino-3-carboxymuconate 6-semialdehyde decarboxylase, H110A tetramer, at pH 8.5 測定日: 2018年7月15日 / 保管温度: 25 °C / セル温度: 25 °C / 照射時間: 1 sec. / フレーム数: 500 / 単位: 1/A /
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距離分布関数 P(R) | ソフトウェア P(R): GNOM 5.0 / ポイント数: 409 /
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結果 | カーブのタイプ: sec /
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